Composition for solubilization of paclitaxel and preparation method thereof

ABSTRACT

The present invention relates to a paclitaxel composition and the preparation methods thereof to solubilize paclitaxel wherein said composition comprises 4˜90% by weight of at least one selected from the monoglycerides, 0.01˜90% by weight of at least one oil and 0.01˜20% by weight of paclitaxel. Also the present invention relates to a paclitaxel composition including emulsifiers and the preparation methods thereof to solubilize paclitaxel wherein said composition comprises 4˜90% by weight of at least one selected from the monoglycerides, 0.01˜90% by weight of at least one oil, 0.01˜90% by weight of at least one emulsifier and 0.01˜20% by weight of paclitaxel. The composition of the present invention is an effective paclitaxel delivery system since the composition solubilizes paclitaxel, does not form aggregates after being dispersed in water, adsorbs well on the intestinal wall, and therefore has high bioavailability.

TECHNICAL FIELD

The present invention relates to a paclitaxel composition and thepreparation methods thereof to solubilize paclitaxel.

BACKGROUND ART

Paclitaxel, an anticancer agent, shows excellent cytotoxicity to variouskinds of cancers such as ovarian cancer, breast cancer, esophaguscancer, melanoma and leukemia. Paclitaxel formulation currently used inclinical remedies has been commercialized in the form of emulsionpreconcentrate (self-emulsifying system) because its water solubility isvery low even compared with an anti-cancer medicine of Bristol-MyersSquibb Company. Taxol® is a commercially available injection agent, inthe form of solution, in which paclitaxel is mixed with solubilizingagent, that is, Cremophor EL (polyoxyethylene 35 castor oil,polyoxylethylaed castor oil and polyoxyethoxylated castor oil) indehydrated alcohol (U.S. Pat. No. 5,438,072). It is known, however, thatthis agent has a limitation in directions and dosage becausesolubilizing agent in Taxol® causes toxic side effects. Therefore, manystudies have been performed to develop new paclitaxel formulations withhigh stability and low toxic effects. There are many patents describinglipid emulsion, polymeric micelles and liposome. In emulsionformulation, not only emulsion using conventional oils and emulsifiersbut also solid lipid nanoparticles, emulsion concentrate and so on havebeen patented. Also other solubilization techniques by utilizingliposome, polymeric nanoparticles and polymeric micelles have beendeveloped. These formulations solubilizing paclitaxel took advantage ofthe accumulated technological advancement already developed for otherinsoluble drugs.

Also, even though paclitaxel is currently used to treat metastaticovarian cancer and breast cancer, it is expected to be prescribed forvarious cancers, especially the metastatic solid tumors (e.g., lungcancer and hepatoma) in the near future. Therefore, market forecast ispromising for paclitaxel.

From the pharmaceutical point of view, Taxol®, the most frequentlyprescribed paclitaxel formulation has a problem of forming precipitationwhen diluted inside the infusion bag due to the low solubility. In-linefilter is used to prevent the precipitation from entering the bloodstream of the patient. The exact dose of paclitaxel, therefore, isunknown and varies from time to time. Also, the plasticizer is known toleak out from the infusion bag made of PVC causing potential healthproblem. From the pharmacological point of view, Cremophor EL, theexcipient can cause severe side-effects such as hypersensitivity,vasodilation, dyspnea, enervation and high blood pressure. From thepharmaceutical and pharmacological points of view, the stability and thesafety of the drug must be improved by developing other administrationroutes and formulations.

The most promising and convenient administration route is considered tobe the oral route. There is a big hurdle to overcome, however, sincepaclitaxel is not absorbed into the body due to the efflux mechanism byp-glycoprotein which exists in the epithelial cell of gastrointestinaltract. Many p-glycoprotein inhibitors are known up to date includingcinchonin, calcium channel blockers such as verapamil anddihydropyridines (for instance nifedipine, nicardipine andnitrendipine), calmodulin antagonist such as trifluoroperazine,antihypertensive such as reserpine, Vinca alkaloids such as vincristineand vinblastine, steroids such as progesterone, antiarrythmics such asamiodarone and quinidine, anthelmintic such as quinacrine and quinine,and immunosuppressants such as cyclosporine A, staurosporine andtacrolimus.

In addition to the increased oral bioavailability of paclitaxel, thep-glycoprotein inhibitors can help overcome multi-drug resistance byinhibiting p-glycoprotein existing in the cancer cells. On the otherhand, paclitaxel is known to be metabolized by hepatic microsomalenzyme. Paclitaxel converts to 6-α-hydroxypaclitaxel and3′-p-hydroxypaclitaxel by CYP2C8 and CYP3A4, respectively.

Cyclosporin A inhibits the formation of 6-α-hydroxypaclitaxel.Doxorubicin, etoposide (VP-16) and cisplatin inhibit the formation of3′-p-hydroxypaclitaxel. And verapamil and tamoxifen inhibit themetabolism of paclitaxel to 6-α-hydroxypaclitaxel and3′-p-hydroxypaclitaxel. Therefore, co-administration of paclitaxel withthe above mentioned metabolism inhibitors could also increase thebioavailability of paclitaxel.

Many formulations have been developed to solubilize paclitaxel. One ofthe most widely used and successful formulations is Taxol®. Lipidemulsion (U.S. Pat. No. 6,391,832 Medical emulsion for lubrication anddelivery of drugs; U.S. Pat. No. 6,348,491 Oil-in-water emulsion forencapsulating paclitaxel) or pre-concentrate which forms transparentcolloidal system in water (U.S. Pat. No. 6,267,985 Clear oil-containingpharmaceutical compositions; U.S. Pat. No. 6,294,192 Triglyceride-freecompositions and methods for improved delivery of hydrophobictherapeutic agents) are also well known. In case of pre-concentrate,clear oil composition forms dispersion of very small particles whoseabsorbance at 400 nm is less than 0.3.

The above formulation is distinctly different from the composition forsolubilization of paclitaxel in the present invention. The compositionin the present invention is coarsely dispersed in water, and theabsorbance at 400 nm is above 0.38 in all cases and between 1 and 4 inmost cases. In other words, efforts have been made to prepare nano-sizedparticles or its pre-concentrate to solubilize paclitaxel in theexisting inventions whereas the current invention discloses thecomposition that does not disperse well and does disperse to formparticles of a few micrometers in size if it does. The merits of thecomposition of the present invention include that paclitaxel does notprecipitate out in the dispersion while it is a high mucoadhesive in theintestine.

One of the main obstacles in commercializing oral paclitaxelformulations is the problem of forming paclitaxel precipitation upondilution with body fluid. Even if the formulation is stable beforedilution, precipitation forms with time in the dispersion. Paclitaxelprecipitation cannot be absorbed into the body in the intestine at all.Once the problem of precipitation formation is solved, however, anotherobstacle, efflux system of p-glycoprotein in the gastrointestinal tract,awaits lowering bioavailability of paclitaxel.

In the present invention, a mucoadhesive lipid, monoolein was used as amain component for oral delivery of paclitaxel. Even though paclitaxelis solubilized in monoolein, it forms precipitation in the cubic phaseof the monoolein/water system. Therefore, we prepared an oilycomposition that does not form paclitaxel precipitation with time evenafter the composition is mixed with water. The cubic phase of themonoolein/water system is composed of ca. 60% (v/v) of water. On theother hand, when more than 10% of oil is added to monoolein, the mixturedoes not form cubic phase, but forms an amorphous composition thatcontains ca. 5˜10% of water. It is worthwhile to note that thiscomposition does not form paclitaxel precipitation. Also, thiscomposition is very mucoadhesive to intestinal wall.

To date, oral paclitaxel formulation that does not requirep-glycoprotein inhibitor has not been developed. Also thebioavailability of the oral paclitaxel formulations was very low evenwhen it is co-administered with p-glycoprotein inhibitor orally.

To overcome the problem of forming paclitaxel precipitation in contactwith water and of low oral bioavailability as mentioned above, thepresent invention provides mucoadhesive compositions for solubilizationof paclitaxel that have high bioavailability when administered alone orwith p-glycoprotein inhibitor and the preparation method thereof.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides compositions for solubilization ofpaclitaxel that is stable for a prolonged period, mucoadhesive, andhighly bioavailable and the preparation method thereof.

More specifically, the present invention provides compositions forsolubilization of paclitaxel including at least one monoglyceride, atleast one oil and paclitaxel and the preparation method thereof.

Also, the present invention provides compositions for solubilization ofpaclitaxel including at least one monoglyceride, at least one oil, atleast one emulsifier and paclitaxel and the preparation method thereof.

In what follows, the present invention will be described in detail.

The present invention provides compositions for solubilization ofpaclitaxel.

Specifically, the above composition is composed of 4˜90% by weight of atleast one selected from the monoglycerides, 0.01˜90% by weight of atleast one oil ad 0.01˜20% by weight of paclitaxel (with respect to thetotal weight of the composition).

The above composition can be prepared by mixing at least onemonoglyceride, at least one oil and paclitaxel at room or elevatedtemperature.

The above monoglycerides are selected from a group consisting of one ormore saturated or unsaturated monoglycerides having 10˜22 carbon atomsin the hydrocarbon chain. Monoglyceride is selected preferably from agroup consisting of monoolein, monopalmitolein, monomyristolein,monoelaidin and monoerucin, and from a group consisting of the mixtureof monoglycerides semi-synthesized from triglycerides of vegetable oranimal oil, and more preferably monoolein.

The above oil is selected preferably from a group consisting oftriglycerides, iodinated oil and vegetable or animal oil that cansolubilize paclitaxel.

The above triglycerides are selected from a group consisting of one ormore saturated or unsaturated triglycerides having 2˜20 carbon atoms inthe hydrocarbon chain. For instance, triacetin, tributyrin, tricaproin,tricaprylin, tricaprin or triolein can be used.

The above iodized oils include iodized poppy seed oil such as Lipiodol,Ethiodol and iodized soybean oil.

The above vegetable oils include soybean oil, cottonseed oil, olive oil,poppyseed oil, linseed oil or sesame oil.

The above animal oils include squalane or squalene.

Also, the above composition can additionally include other additives upto 5% by weight. For instance, the composition can further comprisealcohol, polyol or Cremophor to improve the solubility of paclitaxel,tocopherol or tocopherol acetate to prevent oxidation, fatty acid, fattyacid ester or fatty acid alcohol to increase drug absorption, and otherinsoluble drugs to achieve synergistic effect.

The above insoluble drugs include other anticancer drugs, p-glycoproteininhibitors or hepatic metabolism blockers.

The above other anticancer drugs include doxorubicin, cisplatin,carboplatin, carmustin (BCNU), dacarbazine, etoposide, 5-fluorouracil orpaclitaxel derivatives. The above paclitaxel derivatives includedocetaxel, bromotaxel and taxotere.

The above p-glycoprotein inhibitors include cinchonin, calcium channelblocker, calmodulin antagonist, Vinca alkaloid, antiarrhythmic, steroid,antihypertension drug, anthelmintic or immunosuppressant. The abovecalcium channel blockers include dihydropyridines such as verapamil,nifedipine, nicardipine or nitrendipine. The above calmodulinantagonists include trifluoroperazine. The above antihypertension drugsinclude reserpine. The above Vinca alkaloids include vincristine orvinblastine. The above steroids include progesterone. The aboveantiarrhythmics include amiodarone and quinidine. The aboveanthelmintics include quinacrine and quinine. The aboveimmunosuppressants include cyclosporins, staurosporin and tacrolimus

The above hepatic metabolism blockers include anticancer drugs such ascyclosporin A, doxorubicin, etoposide (VP-16), cisplatin, verapamil andtamoxifen.

The compositions for solubilization of paclitaxel according to thepresent invention can be administered via various routes including oraladministration, buccal administration, mucosal administration, nasaladministration, intraperitoneal administration, subcutaneous injection,intramuscular injection, transdermal administration, intratumoraladministration, and more preferably an oral administration.

The method of preparing the above composition for solubilization ofpaclitaxel comprises the steps of;

-   -   1) preparing the viscous liquid by solubilizing 4˜90% by weight        of at least one monoglyceride compound in 0.01˜90% by weight of        at least one oil by heating to below 50° C. (step 1); and    -   2) preparing homogeneous mixture by solubilizing completely        0.01˜20% by weight of paclitaxel in said mixture in step (1)        (step 2).

The mixture can be heated to 50° C. and sonicated in a bath typesonicator in step (2) to speed up the solubilization process.

Also, the method of preparing the above composition for solubilizationof paclitaxel comprises the steps of;

-   -   1) mixing 4˜90% by weight of at least one monoglyceride        compound, 0.01˜90% by weight of at least one oil and 0.01˜20% by        weight of paclitaxel (step 1); and    -   2) preparing homogeneous mixture by solubilizing completely the        said mixture in step (1) (step 2).

The above mixture can be heated to 50° C. and sonicated in a bath typesonicator or stirred in step (2) to speed up the solubilization process.

The preparation methods described above are only two of many possiblemethods, and other preparation methods can also be used to obtain theabove composition for solubilization of paclitaxel.

Also, the present invention provides the compositions for solubilizationof paclitaxel including emulsifiers.

More particularly, the above composition is composed of 4˜90% by weightof at least one selected from the monoglycerides, 0.01˜90% by weight ofat least one oil, 0.01˜90% by weight of at least one emulsifier and0.01˜20% by weight of paclitaxel (with respect to the total weight ofthe composition).

The above composition can be prepared by adding at least onemonoglyceride, at least one oil, at least one emulsifier and paclitaxelat room or elevated temperature.

The above monoglycerides are selected from a group consisting of one ormore saturated or unsaturated monoglycerides having 10˜22 carbon atomsin the hydrocarbon chain. Monoglyceride is selected preferably from agroup of consisting of monoolein, monopalmitolein, monomyristolein,monoelaidin, and monoerucin, or from a group consisting ofmonoglycerides semi-synthesized from triglycerides of vegetable oranimal oils or their mixture, and more preferably monoolein.

The above oil is selected preferably from a group consisting oftriglycerides, iodinated oil and vegetable or animal oil that cansolubilize paclitaxel.

The above triglycerides are selected from a group consisting of one ormore saturated or unsaturated triglycerides having 2˜20carbon atoms inthe hydrocarbon chain. For instance, triacetin, tributyrin, tricaproin,tricaprylin, tricaprin or triolein can be used.

The above iodized oils include iodized poppy seed oil such as Lipiodol,Ethiodol and iodized soybean oil.

The above vegetable oils include soybean oil, cottonseed oil, olive oil,poppyseed oil, linseed oil or sesame oil.

The above animal oils include squalane or squalene.

The above emulsifier is preferred to select from the group consisting ofa phospholipid, a non-ionic surfactant, an anionic surfactant, acationic surfactant, and bile acid.

The phospholipid is preferred to select from the group consisting of aphosphatidylcholine (PC) and its derivative, a phosphatidylethanolamine(PE) and its derivative, a phosphatidylserine (PS) and its derivative,and a polymeric lipid wherein a hydrophilic polymer is conjugated to thelipid headgroup.

The non-ionic surfactant is selected from the group consisting of apoloxamer (also known as Pluronic: polyoxyethylene-polyoxypropylenecopolymer), a sorbitan ester (Span), a polyoxyethylene sorbitan (Tween)and a polyoxyethylene ether (Brij).

The anionic surfactant is selected from the group consisting of aphosphatidylserine (PS) and its derivative, a phosphatidic acid (PA) andits derivative and sodium dodecyl sulfate (SDS).

The cationic surfactant is selected from the group consisting of1,2-dioleyl-3-trimethylammonium propane (DOTAP),dimethyldioctadecylammonium bromide (DDAB),N-[1-(1,2-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA),1,2-dioleyl-3-ethylphosphocholine (DOEPC) and3β-[N-[(N′,N′-dimethylamino)ethan]carbamoyl]cholesterol (DC-Chol).

The bile acid is selected from the group consisting of cholic acid, itssalt and derivatives; deoxycholic acid, its salt and derivatives;chenocholic acid, its salt and derivatives; and lithocholic acid, itssalt and derivatives.

Other additives can be added to the above to be within 5% by weight. Forinstance, the composition can further comprise alcohol, polyol orCremophor to improve the solubility of paclitaxel, tocopherol ortocopherol acetate to prevent oxidation, and fatty acid, fatty acidester or fatty acid alcohol to increase drug absorption. Depending onthe symptom, other insoluble drug can also be added in the compositionincluding emulsifier according to the present invention.

The above insoluble drugs include other anticancer drugs, p-glycoproteininhibitors or hepatic metabolism blocker.

The above other anticancer drugs include doxorubicin, cisplatin,carboplatin, carmustin (BCNU), dacarbazine, etoposide, 5-fluorouracil orpaclitaxel derivatives. The above paclitaxel derivatives includedocetaxel, bromotaxel and taxotere.

The above p-glycoprotein inhibitors include cinchonin, calcium channelblocker, calmodulin antagonist, Vinca alkaloid, antiarrhythmic, steroid,antihypertension drug, anthelmintic or immunosuppressant. The abovecalcium channel blockers include dihydropyridines such as verapamil,nifedipine, nicardipine or nitrendipine. The above calmodulinantagonists include trifluoroperazine. The above antihypertension drugsinclude reserpine. The above Vinca alkaloids include vincristine orvinblastine. The above steroids include progesterone. The aboveantiarrhythmics include amiodarone and quinidine. The aboveanthelmintics include quinacrine and quinine. The aboveimmunosuppressants include cyclosporins, staurosporin and tacrolimus

The above hepatic metabolism blockers include anticancer drugs such ascyclosporin A, doxorubicin, etoposide (VP-16), cisplatin, verapamil andtamoxifen.

The compositions for solubilization of paclitaxel including emulsifiersaccording to the present invention can be administered via variousroutes including oral administration, buccal administration, mucosaladministration, nasal administration, intraperitoneal administration,subcutaneous injection, intramuscular injection, transdermaladministration, intratumoral administration, and more preferably an oraladministration.

The method of preparing the above composition for solubilization ofpaclitaxel including emulsifiers comprises the steps of;

-   -   1) preparing the viscous liquid by mixing 4˜90% by weight of at        least one monoglyceride compound, 0.01˜90% by weight of at least        one oil, and 0.01˜90% by weight of at least one emulsifier by        heating to below 50° C. (step 1); and    -   2) preparing homogeneous mixture by solubilizing completely        0.01˜20% by weight of paclitaxel in said mixture in step (1)        (step 2).

One of the examples in preparing the composition for solubilization ofpaclitaxel including emulsifiers is as follows. In homogeneous viscousliquid obtained by mixing monoglyceride, oil and emulsifier by heatingto below 50° C., paclitaxel is added. The mixture was stirred orsonicated for 3˜5 minutes at or below 50° C. to obtain homogeneouscomposition.

The method of preparing the above composition for solubilization ofpaclitaxel including emulsifiers can also comprise the steps of;

-   -   1) preparing the paclitaxel solution by solubilizing 0.01˜20% by        weight of paclitaxel in 0.01˜90% by weight of at least one oil        by sonicating in a bath type sonicator (step 1); and    -   2) preparing homogeneous mixture by mixing the paclitaxel        solution in step (1) and 0.01˜90% by weight of at least one        emulsifier and 4˜90% by weight of monoglyceride (step 2).

The preparation methods described above are only two of many possiblemethods, and other preparation method can also be used to obtain theabove composition for solubilization of paclitaxel including emulsifier.

The formulations of the present invention in viscous liquid, gel orsemi-solid form are stable for a long period of time since the physicalproperty of the composition does not change and the components includingpaclitaxel do not degrade with time. Also the compositions forsolubilization of paclitaxel of the present invention is an efficientsolubilization system since they can be easily dispersed in water or inaqueous solutions to become particles bigger than 400 nm in diameter,and the dispersion does not form aggregates with time. Also theabsorbance of the dispersion ranges 1 and 4 at the wavelength of 400 nmindicating that the average particle size is relatively big.

In other words, the composition of the present invention does not formfine dispersion with nano-sized particles, but form coarse dispersionwith the particles of several hundred nanometers to several micrometersin size. The dispersion of the composition of the present invention doesnot form paclitaxel aggregate with time. When administered into thebody, the composition is highly mucoadhesive and adheres onto a widearea of intestinal wall. Paclitaxel in the composition is absorbedthrough the mucosal membrane in the intestine since mucoadhesivemonoolein can be absorbed without further metabolization process.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the total concentration of paclitaxel and itsmetabolites in blood after oral administration of the liquid formulationsolubilizing paclitaxel in Examples 1 and 2 of the present invention.The quantitative analysis of total concentration of paclitaxel and itsmetabolites was performed by ELISA. Taxolφ of Bristol-Myers SquibbCompany was administered via intravenous route as a control group.

-   -   —●—; a group orally administered with liquid formulation for        solubilization of paclitaxel of the present invention (1 mg        paclitaxel, weight ratio of the composition is        monoolein:tricaprylin:paclitaxel=66:33:1, the composition in        Example 1), and    -   —◯—; a group orally administered with liquid formulation for        solubilization of paclitaxel of the present invention (2 mg        paclitaxel, weight ratio of the composition is        monoolein:tricaprylin:paclitaxel=65:33:2, the composition in        Example 2)    -   —▴—; a group intravenously administered with Taxol® of        Bristol-Myers Squibb Company (10 μg paclitaxel).

FIG. 2 is a graph showing the total concentration of paclitaxel and itsmetabolites in blood after oral administration of the liquid formulationcontaining emulsifier with different ratios of paclitaxel. Thequantitative analysis of total concentration of paclitaxel and itsmetabolites was performed by ELISA. Taxol® of Bristol-Myers SquibbCompany was administered orally as a control group.

-   -   —●—; a group orally administered with liquid formulation        containing emulsifier for solubilization of paclitaxel of the        present invention (1 mg paclitaxel, weight ratio of the        composition is monoolein:tricaprylin:Tween        80:paclitaxel=55:28:16:1, the composition in Example 3),    -   —◯—; a group orally administered with liquid formulation        containing emulsifier for solubilization of paclitaxel of the        present invention (1 mg paclitaxel, weight ratio of the        composition is monoolein:tricaprylin:Tween        80:paclitaxel=54:27:16:2, the composition in Example 9),    -   —▴— a group orally administered with liquid formulation        containing emulsifier for solubilization of paclitaxel of the        present invention (1 mg paclitaxel, weight ratio of the        composition is monoolein:tricaprylin:Tween 80:paclitaxel        =55:27:16:3, the composition in Example 10), and    -   —□—; a group orally administered with Taxol® of Bristol-Myers        Squibb Company (1 mg paclitaxel).

FIG. 3 is a graph showing the total concentration of paclitaxel and itsmetabolites in blood after oral administration of the liquid formulationcontaining emulsifier in Example 11 of the present invention. Thequantitative analysis of total concentration of paclitaxel and itsmetabolites was performed by ELISA. Taxol® of Bristol-Myers SquibbCompany was administered intravenously as a control group.

-   -   —●—; a group orally administered with liquid formulation        containing emulsifier for solubilization of paclitaxel of the        present invention (1 mg paclitaxel, weight ratio of the        composition is monoolein:tricaprylin:Pluronic        F68:paclitaxel=55:28:16:1, the composition in Example 11), and    -   —◯—; a group intravenously administered with Taxol® of        Bristol-Myers Squibb company (10 μg paclitaxel).

FIG. 4 is a graph showing the total concentration of paclitaxel and itsmetabolites in blood after oral administration of the liquid formulationcontaining emulsifier in Example 4 of the present invention with orwithout I mg of verapamil. The quantitative analysis of totalconcentration of paclitaxel and its metabolites was performed by ELISA.

-   -   —●—; a group orally administered with liquid formulation        containing emulsifier for solubilization of paclitaxel of the        present invention (1 mg paclitaxel, weight ratio of the        composition is monoolein:tricaprylin:Pluronic        F-68:paclitaxel=55:28:16:1, the composition in Example 4), and    -   —◯—; a group orally administered with verapamil and liquid        formulation containing emulsifier for solubilization of        paclitaxel of the present invention (1 mg paclitaxel, weight        ratio of the composition is monoolein:tricaprylin:Pluronic        F68:paclitaxel=55:28:16:1, the composition in Example 4+1 mg        verapamil).

FIG. 5 is a graph showing the concentration of paclitaxel in blood afteroral administration of the oily composition containing emulsifier(weight ratio of the composition is monoolein:tricaprylin:Tween80:paclitaxel=55:28:16:1). The quantitative analysis of totalconcentration of paclitaxel was performed by HPLC. Taxol® ofBristol-Myers Squibb Company was administered intravenously as a controlgroup.

-   -   —●—; a group orally administered with liquid formulation        containing emulsifier for solubilization of paclitaxel of the        present invention (1 mg paclitaxel), and    -   —◯—; a group intravenously administered with Taxol® of        Bristol-Myers Squibb company (40 μg paclitaxel).

FIG. 6 is a graph showing the total concentration of paclitaxel and itsmetabolites in blood after intraperitoneal or intramuscularadministration of the liquid formulation containing emulsifier in thecomposition of Example 3. The quantitative analysis of totalconcentration of paclitaxel and its metabolites was performed by ELISA.

-   -   —●—; a group intraperitoneally administered with liquid        formulation containing emulsifier for solubilization of        paclitaxel of the present invention (1 mg paclitaxel        monoolein:tricaprylin:Tween 80:paclitaxel=55:28:16:1, the        composition in Example 3),    -   —◯—: a group intramuscularly administered with liquid        formulation containing emulsifier for solubilization of        paclitaxel of the present invention (1 mg paclitaxel, weight        ratio of the composition is monoolein:tricaprylin:Tween        80:paclitaxel=55:28:16:1, the composition in Example 3),

BEST MODE FOR CARRYING OUT THE INVENTION

This invention is explained in more detail based on the followingExamples but they should not be construed as limiting the scope of thisinvention.

Buffer solutions used in the present invention are as follows:

Plate Well Coating Buffer: 50 mM Sodium Phosphate, 0.15 M NaCl and 0.02%sodium azide, pH 7.0 (PBS),

Plate Well Blocking Buffer: PBS containing 1% (w/v) Bovine Serum Albumin(BSA),

Sample Diluting Buffer: PBS containing 0.25% (w/v) BSA, 0.05% (v/v)Tween-20, 20% (v/v) Methanol and 0.02% sodium azide (PBSTM),

Antibody Diluting Buffer: PBS containing 0.25% (w/v) BSA, 0.05% (v/v)Tween-20 and 0.02% sodium azide (PBST),

Wash Buffer: 50 mM Tris HCl, pH 7.0, 0.15 M NaCl, containing 0.05% (v/v)Tween 20 and 0.02% sodium azide (TBST),

Enzyme Substrate Buffer: 25 mM Tris, pH 9.5, 0.15 M NaCl containing 5 mMMgCl2 and 0.02% (w/v) sodium azide.

EXAMPLE 1 Composition for Solubilization of Paclitaxel According to theChange in the Composition Ratio (1)

(1)-Manufacturing Composition for Solubilization of Paclitaxel

Viscous oily solution was prepared by mixing 1 g monoolein and 0.5 gtricaprylin and warmed at 40° C. Fifteen milligrams of paclitaxel wasadded into the oily solution and sonicated in a bath type sonicator forcomplete solubilization to obtain a liquid formulation.

(2) Property Analysis of Thus Prepared Composition for Solubilization ofPaclitaxel

The size of the emulsion particles were measured by using MalvernZetasizer (Malvern Instruments Limited, England) after diluting theemulsion by adding 3 mL of distilled water with 2 μL of thus obtainedliquid formulation. An average particle size and polydispersity wasobtained by measuring a given formulation three times (Orr, Encyclopediaof emulsion technology, 1, 369-404, 1985). The polydispersity wasobtained as the variance indicated by the logarithmic scale in thelogarithmic normal distribution function. This method was used inmeasuring the particle size and the polydispersity throughout thefollowing Examples.

The above composition exists as semi-solid or solid at room temperatureand in the refrigerator, respectively, but as liquid at or above 40° C.Paclitaxel precipitation was not observed under polarized lightmicroscope, and phase separation was not observed either. Unstabledispersion with the average particle size of 1230 nm was obtained whenthe above composition was vortexed for 10 s in water. Paclitaxelprecipitation was not observed under polarized light microscope 24 hourafter preparing the dispersion.

EXAMPLE 2 Composition for Solubilization of Paclitaxel According to theChange in the Composition Ratio (2)

Viscous oily solution was prepared by mixing 1 g monoolein and 0.5 gtricaprylin and warmed at 40° C. Thirty milligrams of paclitaxel wasadded into the oily solution and sonicated in a bath type sonicator forcomplete solubilization. The above composition exists as semi-solid orsolid at room temperature and in the refrigerator, respectively, but asliquid at or above 40° C. Paclitaxel precipitation was not observedunder polarized light microscope, and phase separation was not observedeither. Unstable dispersion with the average particle size of 2080 nmwas obtained when the above composition was vortexed for 10 s in water.Paclitaxel precipitation was not observed under polarized lightmicroscope 24 hour after preparing the dispersion.

The results of the Examples 1 and 2 are summarized in the followingTable 1. TABLE 1 Content (weight %) Particle size (nm) MonooleinTricaprylin Paclitaxel (polydispersity) Example 66 33 1 1230 (0.200) 165 33 2 2080 (1.000) 2

EXAMPLE 3 Composition for Solubilization of Paclitaxel IncludingEmulsifiers According to the Change in the Composition Ratio (1)

Viscous oily solution was prepared by mixing completely 1 g monoolein,0.5 g tricaprylin and 0.3 9 of Tween 80, and warmed at 40° C. Eighteenmilligrams of paclitaxel was added into the oily solution and sonicatedin a bath type sonicator for complete solubilization. Dispersion withthe average particle size of 600 nm was obtained when the abovecomposition was vortexed for 10 s in water. Paclitaxel precipitation wasnot observed under polarized light microscope 24 hour after preparingthe dispersion. The above composition exists as semi-solid or solid atroom temperature and in the refrigerator, respectively, but as liquid ator above 40° C.

EXAMPLE 4 Composition for Solubilization of Paclitaxel IncludingEmulsifiers According to the Change in the Composition Ratio (2)

The composition and dispersed liquid were prepared the same as those ofthe Example 1 with the exception that 1 g monoolein, 1 g tricaprylin,0.4 g of Tween 80 and 10 mg of paclitaxel were used, and their particlesize and polydispersity were measured by the same methods in theExample 1. Dispersion with the average particle size of 530 nm wasobtained. Paclitaxel precipitation was not observed under polarizedlight microscope, and phase separation was not observed either. Theabove composition exists as semi-solid or solid at room temperature andin the refrigerator, respectively, but as liquid at or above 40° C.

The results of the Examples 3 and 4 are summarized in the followingTable 2. TABLE 2 Content (weight %) Mono- Tricapry- Tween Particle size(nm) olein lin 80 Paclitaxel (polydispersity) Example 55 28 16 1 600(0.200) 3 41.5 41.5 16.6 0.4 530 (1.000) 4

COMPARATIVE EXAMPLE 1 Composition Including Emulsifier forSolubilization of Paclitaxel Without Oil (1)

The composition and dispersed liquid were prepared the same as those ofthe Example 1 with the exception that 1 g monoolein, 0.2 g of Tween 80and 12 mg of paclitaxel were used, and their particle size andpolydispersity were measured by the same methods in the Example 1.Dispersion with the average particle size of 700 nm was obtained.Paclitaxel precipitation was observed under polarized light microscope,and the dispersion became unstable 1 hour after preparation.

COMPARATIVE EXAMPLE 2 Composition Including Emulsifier forSolubilization of Paclitaxel Without Oil (2)

The composition and dispersed liquid were prepared the same as those ofthe Example 1 with the exception that 1 g monoolein, 0.24 g of PluronicF-68 and 12.4 mg of paclitaxel were used, and their particle size andpolydispersity were measured by the same methods in the Example 1.Dispersion with the average particle size of 640 nm was obtained.Paclitaxel precipitation was observed under polarized light microscope,and the dispersion became unstable 1 hour after preparation.

COMPARATIVE EXAMPLE 3 Composition Including Emulsifier forSolubilization of Paclitaxel Without Monoolein (1)

The composition and dispersed liquid were prepared the same as those ofthe Example 1 with the exception that 1 g tricaprylin, 0.2 g of Tween 80and 12 mg of paclitaxel were used, and their particle size andpolydispersity were measured by the same methods in the Example 1.Dispersion with the average particle size of 560 nm was obtained.Paclitaxel precipitation was not observed under polarized lightmicroscope, and phase separation was not observed either.

EXAMPLE 5 Composition for Solubilization of Paclitaxel IncludingEmulsifiers According to the Change in the Oil (1)

The composition and dispersed liquid were prepared the same as those ofthe Example 1 with the exception that 1 g monoolein, 0.5 g tributyrin,0.3 g of Tween 80 and 18 mg of paclitaxel were used and their particlesize and polydispersity were measured by the same methods in theExample 1. Dispersion with the average particle size of 950 nm wasobtained. Paclitaxel precipitation was not observed under polarizedlight microscope, and phase separation was not observed either, 24 hoursafter preparing the dispersion. The above composition exists assemi-solid or solid at room temperature and in the refrigerator,respectively, but as liquid at or above 40° C.

EXAMPLE 6 Composition for Solubilization of Paclitaxel IncludingEmulsifiers According to the Change in the Oil (2)

The composition and dispersed liquid were prepared the same as those ofthe Example 1 with the exception that 1 g monoolein, 0.5 g lipiodol(Lipiodol Ultra-fluid, Laboratoire Guerbet, France, Iodine content: 38%by weight), 0.3 g of Tween 80 and 18 mg of paclitaxel were used andtheir particle size and polydispersity were measured by the same methodsin the Example 1. Dispersion with the average particle size of 680 nmwas obtained. Paclitaxel precipitation was not observed under polarizedlight microscope, and phase separation was not observed either, 24 hoursafter preparing the dispersion. The above composition exists assemi-solid or solid at room temperature and in the refrigerator,respectively, but as liquid at or above 40° C.

EXAMPLE 7 Composition for Solubilization of Paclitaxel IncludingEmulsifiers According to the Change in the Oil (3)

The composition and dispersed liquid were prepared the same as those ofthe Example I with the exception that 1 g monoolein, 0.5 g squalane(Sigma Chemical Company), 0.3 g of Tween 80 and 18 mg of paclitaxel wereused and their particle size and polydispersity were measured by thesame methods in the Example 1. Dispersion with the average particle sizeof 598 nm was obtained. Paclitaxel precipitation was not observed underpolarized light microscope, and phase separation was not observedeither, 24 hours after preparing the dispersion. The above compositionexists as semi-solid or solid at room temperature and in therefrigerator, respectively, but as liquid at or above 40° C.

EXAMPLE 8 Composition for Solubilization of Paclitaxel IncludingEmulsifiers According to the Change in the Oil (4)

The composition and dispersed liquid were prepared the same as those ofthe Example 1 with the exception that 1 g monoolein, 0.5 g safflowerseed oil (Sigma Chemical Company), 0.3 g of Tween 80 and 18 mg ofpaclitaxel were used and their particle size and polydispersity weremeasured by the same methods in the Example 1. Dispersion with theaverage particle size of 1040 nm was obtained. Paclitaxel precipitationwas not observed under polarized light microscope, and phase separationwas not observed either, 24 hours after preparing the dispersion. Theabove composition exists as semi-solid or solid at room temperature andin the refrigerator, respectively, but as liquid at or above 40° C.

The results of the Examples 5-8 are summarized in the following Table 3.TABLE 3 Particle size (nm) Oil (polydispersity) Example Tributyrin  950(0.661) 5 Lipiodol  680 (1.000) 6 Squalane  597 (0.550) 7 Safflower seedoil 1040 (0.497) 8*Monoolein:Oil:Tween 80:Paclitaxel = 55:28:16:1 (Weight ratio)

EXAMPLE 9 Composition for Solubilization of Paclitaxel IncludingEmulsifiers According to the Change in the Paclitaxel Content (1)

The composition and dispersed liquid were prepared the same as those ofthe Example 1 with the exception that 1 g monoolein, 0.5 g tricaprylin,0.3 g of Tween 80 and 38 mg of paclitaxel were used and their particlesize and polydispersity were measured by the same methods in theExample 1. Dispersion with the average particle size of 1450 nm wasobtained. Paclitaxel precipitation was not observed under polarizedlight microscope, and phase separation was not observed either, 24 hoursafter preparing the dispersion. The above composition exists assemi-solid or solid at room temperature and in the refrigerator,respectively, but as liquid at or above 40° C.

EXAMPLE 10 Composition for Solubilization of Paclitaxel IncludingEmulsifiers According to the Change in the Paclitaxel Content (2)

The composition and dispersed liquid were prepared the same as those ofthe Example 1 with the exception that 19 monoolein, 0.5 g tricaprylin,0.3 g of Tween 80 and 54 mg of paclitaxel were used and their particlesize and polydispersity were measured by the same methods in theExample 1. Dispersion with the average particle size of 1630 nm wasobtained. Paclitaxel precipitation was not observed under polarizedlight microscope, and phase separation was not observed, 24 hours afterpreparing the dispersion. Unlike other compositions in Examples 1˜7, theabove composition exists as liquid or solid at room temperature and inthe refrigerator, respectively.

The results of the Examples 9 and 10 are summarized in the followingTable 4. TABLE 4 Content (weight %) Mono- Tricapry- Tween Particle size(nm) olein lin 80 Paclitaxel (polydispersity) Example 55 27 16 2 1450(1.000)  9 54 27 16 3 1630 (1.000) 10

EXAMPLE 11 Composition for Solubilization of Paclitaxel IncludingEmulsifiers According to the change in the Emulsifier

The composition and dispersed liquid were prepared the same as those ofthe Example 3 with the exception that Pluronic F68 (BASF Company) wasused instead of Tween 80. Dispersion with the average particle size of420 nm (polydispersity 0.284) was obtained. Paclitaxel precipitation wasnot observed under polarized light microscope, and phase separation wasnot observed either, 24 hours after preparing the dispersion. The abovecomposition exists as semi-solid or solid at room temperature and in therefrigerator, respectively, but as liquid at or above 40° C.

EXAMPLE 12 In vivo Oral Administration of Composition for Solubilizationof Paclitaxel (1)

Animal experiments were performed by using the composition for thesolubilization of paclitaxel prepared in the above Example 1.

{circle around (1)} Oral Administration of Composition for theSolubilization of Paclitaxel

One hundred microliters of the mucoadhesive formulation containing 1 mgpaclitaxel was administered into Balb/C mouse (6˜7 weeks old, female)fasted for 4 hours previously by using a gastric sonde. As a controlgroup, 167 μl of Taxol® of Bristol-Myers Squibb Company dispersed in 0.5ml of water (corresponding to 1 mg of paclitaxel) was administeredorally. One hundred microliters of tricaprylin/Tween 80 compositionwithout monoolein (containing 1 mg paclitaxel) prepared in ComparativeExample 3 was used as another control group and administered orally.One, 2, 3 and 4 h after the oral administration of the compositions, theconcentration of paclitaxel in the blood collected from the eye wasdetermined.

As a control for oral administration, Taxol® of Bristol-Myers SquibbCompany was administered intravenously into Balb/C mouse (6˜7 weeks old,female), and the concentration of paclitaxel in blood was determined for4 hours after intravenous injection. After dispersing 0.1 ml of Taxol®in 5.9 ml water, 0.1 ml of the dispersion (corresponding to 10 μg ofpaclitaxel) was administered by bolus injection into the tail vein ofBalb/C mouse fasted for 4 hours. Blood was collected 0.5, 1, 2 and 4 hafter the administration of the compositions, and the concentration ofpaclitaxel in the blood collected from the eye was determined.

{circle around (2)} Determination of Total Concentration of Paclitaxeland its Metabolites in Blood (ELISA Method)

The total concentration of paclitaxel and its metabolites in blood wasdetermined by using Anti-taxane monoclonal kit (Model number 8A10) fromHawaii Biotech Company. Paclitaxel is known to be converted to6-α-hydroxypaclitaxel and 3′-p-hydroxypaclitaxel by CYP2C8 and CYP3A4,respectively. Various metabolites including the primary metabolites ofpaclitaxel exist in the blood. Anti-taxane monoclonal kit enables us todetermine the concentration of paclitaxel and all of the metabolitescontaining taxane ring (Grothaus, G. P., Bignami, G. S., O'Malley, S.,Harada, K. E., Byrnes, J. B., Waller, D. F., Raybould, T. J. G.,Mcguire, M. T. and Alvaro, B., Taxane-specific monoclonal antibodies:measurement of Taxol, baccatin III, and ‘total taxanes’ in Taxusbrevifolia extracts by enzyme immunoassay. J. Nat. Prod. 58, pp.1003-1014, 1995).

The blood sample was serially diluted 4 times. Taxol-protein coatingantigen (blue label) was diluted 100 times by phosphate buffered saline(PBS). After 100 μl of the diluted antigen solution was put into eachwell of the 96-well plate, the plate was incubated for 1 hour. After theplate was washed 4 times with TBST, it was blocked by adding PBScontaining 1% bovine serum albumin for 1 hour. After each well waswashed continuously four times with TBST, 50 μl of the serially dilutedsamples were put into each well. After diluting HBC Taxol Standard (REDlabel) serially with PBST, 50 μl of the diluted standard solution wasput into each well. Fifty microliters of the antibody solution preparedby mixing 4.5 ml PBST and 50 μl of anti-taxane rabbit antibody (greenlabel) was added in each well. After the wells were washed four timeswith TBST, 100 μl of secondary antibody solution diluted 1000 times withPBST was added and incubated for one hour. After washing the wells fourtimes with TBST, 200 μl of pNPP solution at 1 mg/ml was added in eachwell. After incubating the plate for 1 hour at room temperature, theabsorbance was measured at 414 nm by ELISA reader and compared with thatat 690 nm for quantitative analysis.

{circle around (3)} Results of Oral Administration of Composition forthe Solubilization of Paclitaxel

The changes in the paclitaxel concentration in blood with time are shownin FIG. 1. When the bioavailability of paclitaxel upon bolus injectionwas set to 100%, the relative bioavailability upon oral administrationof composition for the solubilization of paclitaxel was calculated bythe following formula.${{Bioavailability}(\%)} = {\frac{{AUC}\quad{oral}}{{AUC}\quad{iv}} \times \frac{{DOSE}\quad{iv}}{{DOSE}\quad{oral}} \times 100}$

Wherein, AUCoral and AUCiv represent area under the curve after oral andintravenous administration, respectively, and DOSEiv and DOSEoralrepresent the paclitaxel dose for the oral and intravenousadministration, respectively. The bioavailability upon oraladministration of composition for the solubilization of paclitaxel whencompared to the bolus injection was 19.5%.

EXAMPLE 13 In vivo Oral Administration of Composition for Solubilizationof Paclitaxel (2)

Fifty microliters of the composition for solubilizing paclitaxelincluding emulsifier prepared in Example 2 containing 1 mg paclitaxelwas administered into Balb/C mouse as in Example 12. As a control fororal administration, Taxol® of Bristol-Myers Squibb Company wasadministered intravenously (corresponding to 10 μg of paclitaxel/mouse).The total concentrations of paclitaxel and its metabolites in blood withtime are determined by ELISA. FIG. 1 shows the result in comparison tothat of Example 12. The bioavailability upon oral administration ofcomposition for the solubilization of paclitaxel when compared to thebolus injection was 9.5±5%.

EXAMPLE 14 In vivo Oral Administration of Composition for Solubilizationof Paclitaxel Including Emulsifier—Effect of Paclitaxel Content

The compositions for solubilizing paclitaxel including emulsifierprepared in Examples 3, 9 and 10 containing 1 mg paclitaxel wereadministered into Balb/C mouse by using identical method as in Example12. As control groups, Taxol® of Bristol-Myers Squibb Company wasadministered orally (corresponding to 1 mg of paclitaxel) as well asintravenously (corresponding to 10 μg/mouse) as in Example 11. The totalconcentrations of paclitaxel and its metabolites in blood with time aredetermined by ELISA as shown in FIG. 2. The bioavailability calculatedby setting that of Taxol® upon bolus injection in Example 12 to 100% islisted in Table 5. The bioavailability upon oral administration ofcomposition for the solubilization of paclitaxel including emulsifierwhen compared to the bolus injection was approximately 10˜30%. On theother hand, the bioavailability of orally administered Taxol® was1.7±3%. TABLE 5 Oral formulation Content (weight %) Bioavail- MonooleinTricaprylin Tween 80 Paclitaxel ability (%) Example 55 28 16 1 29.5  355 27 16 2 12.1  9 54 27 16 3 11.6 10

EXAMPLE 15 In vivo Oral Administration of Composition for Solubilizationof Paclitaxel Including Emulsifier—Effect of Emulsifier

The compositions for solubilizing paclitaxel including emulsifierprepared in Example 11 containing 1 mg paclitaxel were administered intoBalb/C mouse by using identical method as in Example 12. As a controlgroup, Taxol® of Bristol-Myers Squibb Company was administeredintravenously (corresponding to 10 μg/mouse). The total concentrationsof paclitaxel and its metabolites in blood with time are determined byELISA as shown in FIG. 3. The bioavailability upon oral administrationof composition for the solubilization of paclitaxel including emulsifierwhen compared to the bolus injection was approximately 13.4±4%.

EXAMPLE 16 In vivo Oral Administration of Composition for Solubilizationof Paclitaxel Including Emulsifier—Coadministration of Verapamil

The composition for solubilizing paclitaxel including emulsifierprepared in Example 4 containing 1 mg paclitaxel were administered intoBalb/C mouse by using identical method as in Example 12. Another groupof mice were administered with the composition for solubilizingpaclitaxel including emulsifier prepared in Example 4 containing 1 mgpaclitaxel and 1 mg of verapamil. The total concentrations of paclitaxeland its metabolites in blood with time are determined by ELISA as shownin FIG. 4. The bioavailability upon oral administration of compositionfor the solubilization of paclitaxel including emulsifier when comparedto the bolus injection of Taxol® in Example 12 was approximately 2.7%.When verapamil was co-administered, however, the oral bioavailabilityincreased to 17.6%.

EXAMPLE 17 In vivo Oral Administration of Composition for Solubilizationof Paclitaxel Including Emulsifier—Determination of PaclitaxelConcentration by HPLC

The composition for solubilizing paclitaxel including emulsifierprepared in Example 3 containing 1 mg paclitaxel were administered intoBalb/C mouse by using identical method as in Example 12. Paclitaxelconcentration was determined by HPLC. When the above ELISA method wasused, total concentration of paclitaxel as well as its metabolites isdetermined. HPLC analysis, however, detects intact paclitaxel moleculeonly in the sample.

{circle around (1)} Determination of paclitaxel concentration in blood(HPLC method)

Plasma was obtained by centrifuging collected blood for 10 min at 3000rpm and stored at −20° C. until being analyzed. Into 200 μl of plasma,10 μl of butyl-p-hydroxybenzoic acid solution dissolved in acetonitrilat 100 μg/ml was added as an internal standard. The sample was extractedby adding 1 ml tert-butylmethylether and vortexed for 30 seconds. Toseparate the organic layer, the above solution was centrifuged for 5 minat 3000 rpm, and 0.8 ml of the organic layer was taken out and driedunder the stream of nitrogen atmosphere. The residue was dissolved in 80μl of 60% acetonitril. Thirty microliters of the above sample wasinjected into HPLC to analyze the concentration of paclitaxel. HPLCsystem consists of Shisheido Nanospace Semimicro Column HPLC, SI-1/2001pump, SI-1/2002 UV-VIS detector, SI-1/2004 column oven, SI-1/2003autosampler, SI-1/2009 degassing unit, SI-1/2011, 2012 rotary highpressure 6-way valve and Shisheido-MicroChrom software. CAPCELPAK MFPh-2 cartridge (4.6×20 mm) was used as pre-treatment column, andseparation of compounds was achieved using a CAPCELL PAK C18 UG120(2.0×250 mm) concentrating column. Mobile phase was 50% phosphate buffersolution (0.1%, pH=6.86) (pump A) and 50% acetonitril (pump B), andcolumn temperature was maintained at 30° C. Paclitaxel concentration inblood was analyzed by column switching method. In case of pre-treatmentcolumn and analytical column, the flow rates of the mobile phase, 0.1%phosphate buffer/acetonitril solution (84:16 by weight) were 0.5 ml/minand 0.1 ml/min, respectively.

The switching valve was set to A position to dispose of the eluent ofthe pre-treatment column, and the eluent of the analytical column wasallowed to go through the UV detector. Since the internal standard andpaclitaxel were detected at 4.1˜7.2 min and 6.4˜9.3 min, respectively,after going through the pre-treatment column, switching valve was set toB position at 4.0 min to allow the eluent of the pre-treatment column togo through the concentrating column. At 10 min, the switching valve wasset to A position again, to change the flow from the concentratingcolumn to analytical column. The chromatograms were collected at 227 nmand analyzed by using Syscon software (Shisheido).

{circle around (2)} Oral Administration of Composition for theSolubilization of Paclitaxel Including Emulsifier

The composition for solubilizing paclitaxel including emulsifierprepared in Example 3 containing 1 mg paclitaxel were administered intoBalb/C mouse by using identical method as in Example 12. As a controlfor oral administration, Taxol® of Bristol-Myers Squibb Company wasadministered intravenously into Balb/C mouse (6˜7 weeks old, female),and the concentration of paclitaxel in blood was determined for 4 hours.After dispersing 0.4 ml of Taxol® in 5.6 ml water, 0.1 ml of thedispersion (corresponding to 40 μg of paclitaxel) was administered bybolus injection into the tail vein of Balb/C mouse fasted for 4 hours.Blood was collected from the eye up to 4 h after the administration ofthe compositions, the concentration of paclitaxel in the collected bloodwas determined. Paclitaxel concentration was determined by HPLC as shownin FIG. 5. The bioavailability upon oral administration of compositionfor the solubilization of paclitaxel including emulsifier when comparedto the bolus injection was approximately 30.3%.

EXAMPLE 18 In vivo Intraperitoneal Administration of Composition forSolubilization of Paclitaxel Including Emulsifier

Fifty microliters of the composition for solubilizing paclitaxelincluding emulsifier prepared in Example 3 containing 1 mg paclitaxelwas administered into Balb/C mouse via intraperitoneal administration.The total concentrations of paclitaxel and its metabolites in blood withtime are determined by ELISA as shown in FIG. 6.

EXAMPLE 19 In vivo Intramuscular Administration of Composition forSolubilization of Paclitaxel Including Emulsifier

Fifty microliters of the composition for solubilizing paclitaxelincluding emulsifier prepared in Example 3 containing 1 mg paclitaxelwere administered into Balb/C mouse via intramuscular administration.The total concentrations of paclitaxel and its metabolites in blood withtime are determined by ELISA as shown in FIG. 6.

INDUSTRIAL APPLICABILITY

As described above, the composition for solubilizing paclitaxelaccording to the present invention can solubilize paclitaxel stably andalso does not form precipitates of paclitaxel when dispersed in water.Also in aqueous environment, the composition forms unstable dispersionof particles of 30 nm˜5 μm. Paclitaxel concentration in blood can bemaintained for more than 4 hours when the composition for solubilizingpaclitaxel according to the present invention is delivered via oral,intraperitoneal and intramuscular administration.

1. A composition for solubilization of paclitaxel comprising 4˜90% by weight of at least one monoolein, 0.01˜90% by weight of an oil chosen from triglyceride, iodized oil, vegetable oil and animal oil, and 0.01˜20% by weight of paclitaxel so that the ratio of monoolein to oil is more than 1:1. 2.-4. (canceled)
 5. The composition for solubilization of paclitaxel according to claim 1, wherein said triglyceride is chosen from saturated and unsaturated triglycerides having 2˜20 carbon atoms in each hydrocarbon chain.
 6. The composition for solubilization of paclitaxel according to claim 1, wherein said triglyceride is chosen from triacetin, tributyrin, tricaproin, tricaprylin, tricaprin and triolein; wherein said iodized oil is chosen from Lipiodol, iodized poppy seed oil, Ethiodol and iodized soybean oil: wherein said vegetable oil is chosen from soybean oil, cottonseed oil, olive oil, poppyseed oil, linseed oil and sesame oil; and wherein said animal oil is chosen from squalane and squalene. 7.-9. (canceled)
 10. The composition for solubilization of paclitaxel according to claim 1 additionally comprising 0.01˜5% by weight of an additive.
 11. The composition for solubilization of paclitaxel according to claim 10, wherein the additive is chosen from Cremophor, tocopherol, tocopherol acetate, a fatty acid acids a fatty acid ester, a fatty acid alcohol, an insoluble drug, an alcohol and a polyol.
 12. The composition for solubilization of paclitaxel according to claim 11, wherein the insoluble drug is chosen from an anticancer drug, a p-glycoprotein inhibitor and a hepatic metabolism blocker; wherein the alcohol is chosen from methanol, ethanol, propanol and isopropanol; and wherein to polyol is chosen from ethyleneglycol, propyleneglycol and polyethyleneglycol.
 13. The composition for solubilization of paclitaxel according to claim 12, wherein the anticancer drug is chosen from doxorubicin, cisplatin, carboplatin, carmustin (BCNU), dacarbazine, etoposide, 5-fluorouracil and a paclitaxel derivative chosen from docetaxel, bromotaxel and taxotere: wherein said p-glycoprotein inhibitor is chosen from cinchonin, a calcium channel blocker, a calmodulin antagonist, an antihypertensive, a Vinca alkaloid, a steroid, an antiarrhythmic, an anthelmintic and an immunosuppressant, and wherein said hepatic metabolism blocker is chosen from an anticancer drug chosen from cyclosporin A, doxorubicin, etoposide (VP-16) and cisplatin, verapamil and tamoxifen. 14.-15. (canceled)
 16. The composition for solubilization of paclitaxel according to claim 13, wherein the calcium channel blocker is chosen from verapamil and a dihydropyridine chosen from nifedipine, nicardipine and nitrendipine; wherein the calmodulin antagonist is chosen from trifluoroperazine; wherein the antihypertensive is reserpine; wherein the Vinca alkaloid is chosen from vincristine and vinblastine; wherein the steroid is progesterone: wherein the antiarrhythmic is chosen from amiodarone and quinidine; wherein the anthelmintic is chosen from quinacrine and quinine; and wherein the immunosuppressant is chosen from cyclosporine A, staurosporine and tacrolimus. 17.-26. (canceled)
 27. The composition for solubilization of paclitaxel according to claims 1, wherein the administration route is chosen from oral administration, buccal administration, mucosal administration, intranasal administration, intraperitoneal administration, subcutaneous injection, intramuscular injection, transdermal administration, intratumoral injection.
 28. A method of preparing the composition for solubilization of paclitaxel according to claims 1, wherein said method comprises the steps of: (1) solubilizing 4˜90% by weight of at least one menoglycride compound monoolein in 0.01˜90% by weight of an oil chosen from triglyceride, iodized oil, vegetable oil and animal oil so that the ratio of monoolein to oil is more than 1:1; and (2) solubilizing completely 0.01˜20% by weight of paclitaxel in said mixture in step (1) by stirring.
 29. The preparation method according to claim 28, wherein the said mixture is heated to 50° C. in step (1) to speed up the solubilization process.
 30. The preparation method according to claim 28, wherein the said mixture is heated to 50° C. and sonicated in a bath type sonicator in step (2) to speed up the solubilization process.
 31. A method of preparing the composition for solubilization of paclitaxel according to claims 1, wherein said method comprises the steps of mixing 4˜90% by weight of at least one moneglyccride compound monoolein, 0.01˜90% by weight of an oil chosen from triglyceride, iodized oil, vegetable oil and animal oil and 0.01˜20% by weight of paclitaxel so that the ratio of monoolein to oil is more than 1:1 and solubilizing completely.
 32. The preparation method according to claim 31, wherein the said mixture is heated to 50° C. and sonicated in a bath type sonicator to speed up the solubilization process.
 33. A composition for solubilization of paclitaxel including emulsifier comprising 4˜90% by weight of at least one monoolein, 0.01˜90% by weight an oil chosen from triglyceride, iodized oil, vegetable oil and animal oil, 0.01˜90% by weight of at least one emulsifier and 0.01˜20% by weight of paclitaxel so that the ratio of monoolein to oil is more than 1:1. 34.-36. (canceled)
 37. The composition for solubilization of paclitaxel including emulsifier according to claim 33, wherein said triglyceride is chosen from saturated and unsaturated triglycerides having 2˜20 carbon atoms in each hydrocarbon chain.
 38. The composition for solubilization of paclitaxel including emulsifier according to claim 33, wherein said triglyceride is chosen from triacetin, tributyrin, tricaproin, tricaprylin, tricaprin and triolein; wherein said iodized oil is chosen from Lipiodol, iodized poppy seed oil, Ethiodol and iodized soybean oil; wherein said vegetable oil is chosen from soybean oil, cottonseed oil, olive oil, poppyseed oil, linseed oil and sesame oil; and wherein said animal oil is chosen from squalane and squalene. 39.-41. (canceled)
 42. The composition for solubilization of paclitaxel including emulsifier according to claim 33, wherein said emulsifier is chosen from a phospholipid, a non-ionic surfactant, an anionic surfactant, a cationic surfactant and bile acid.
 43. The composition for solubilization of paclitaxel including emulsifier according to claim 42, wherein said phospholipid is chosen from a phosphatidylcholine (PC) and its derivative, a phosphatidylethanolamine (PE) and its derivative, a phosphatidylserine (PS) and its derivative, and a polymeric lipid wherein a hydrophilic polymer is conjugated to the lipid headgroup; wherein said non-ionic surfactant is chosen from a poloxamer (Pluronic: polyoxyethylene-polyoxypropylene copolymer), a sorbitan ester (sorbitan esters; Span), a polyoxyethylene sorbitan (Tween) and a polyoxyethylene ether (Brij); wherein said anionic surfactant is chosen from a phosphatidylserine (PS) and its derivative, a phosphatidic acid (PA) and its derivative, and sodium dodecyl sulfate (SDS): wherein said cationic surfactant is chosen from 1,2-dioleyl-3-trimethylammonium propane (DOTAP) dimethyldioctadecylammonium bromide (DDAB), N-[1-(1,2-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA), 1,2-dioleyl-3-ethylphosphocholic acid (DOEPC) and 3β[-N-[(N′,N′-dimethylamino)ethan]carbamoyl]cholesterol (DC-Chol); and wherein said bile acid is chosen from cholic acid its salt and derivatives; deoxycholic acid, its salt and derivatives; chenocholic acid, its salt and derivatives; and lithocholic acid, its salt and derivatives. 44.-47. (canceled)
 48. The composition for solubilization of paclitaxel including emulsifier according to claim 33 additionally comprising 0.01˜5% by weight of other additives.
 49. The composition for solubilization of paclitaxel including emulsifier according to claim 48, wherein said other additives are chosen from Cremophor, tocopherol, tocopherol acetate, a fatty acid, a fatty acid ester, a fatty acid alcohol, an insoluble drug, an alcohol and a polyol.
 50. The composition for solubilization of paclitaxel including emulsifier according to claim 49, wherein said insoluble drugs are chosen from an anticancer drug, a p-glycoprotein inhibitor and a hepatic metabolism blocker; wherein the alcohol is chosen from methanol, ethanol, propanol and isopropanol; and wherein the polyol is chosen from ethyleneglycol, propyleneglycol and polyethyleneglycol.
 51. The composition for solubilization of paclitaxel including emulsifier according to claim 50, wherein the anticancer drug is chosen from doxorubicin, cisplatin, carboplatin, carmustin (BCNU), dacarbazine, etoposide, 5-fluorouracil and paclitaxel derivatives wherein the paclitaxel derivative is chosen from docetaxel, bromotaxel and taxotere; wherein the p-glycoprotein inhibitor is chosen from cinchonins, calcium channel blockers, calmodulin antagonists, Vinca alkaloids, antiarrhythmics, steroids, antihypertension drugs, anthelmintics and immunosuppressants; and wherein the hepatic metabolism blocker is chosen from a anticancer drug chosen from cyclosporin A, doxorubicin, etoposide (VP-16) and cisplatin, verapamil and tamoxifen. 52.-53. (canceled)
 54. The composition for solubilization of paclitaxel including emulsifier according to claim 51, wherein the calcium channel blocker is a dihydropyridine chosen from verapamil, nifedipine, nicardipine and nitrendipine; wherein said calmodulin antagonist is trifluoroperazine; wherein the antihypertension drug is reserpine; wherein the Vinca alkaloid is chosen from vincristine and vinblastine; wherein the steroid is progesterone: wherein the antiarrhythmic is chosen from amiodarone and quinidine; wherein the anthelmintic is chosen from quinacrine and quinine; and wherein the immunosuppressant is chosen from cyclosporins, staurosporin and tacrolimus. 55.-64. (canceled)
 65. The composition for solubilization of paclitaxel including emulsifier according to claims 33, wherein the administration route is chosen from oral administration, buccal administration, mucosal administration, intranasal administration, intraperitoneal administration, subcutaneous injection, intramuscular injection, transdermal administration and intratumoral injection.
 66. A method of preparing the composition for solubilization of paclitaxel including emulsifier according to claims 33, wherein said method comprises the steps of: (1) preparing the viscous liquid by mixing 4˜90% by weight of monoolein, 0.01˜90% by weight of an oil chosen from triglyceride, iodized oil, vegetable oil and animal oil and 0.01˜90% by weight of emulsifier so that the ratio of monoolein to oil is more than 1:1 by heating to below 50° C. (step 1): and (2) preparing homogeneous mixture by solubilizing completely 0.01˜20% by weight of paclitaxel in said mixture in step (1) (step 2).
 67. The method of preparing the composition for solubilization of paclitaxel including emulsifier according to claim 66, wherein the said mixture is heated to 50° C. in step (1) to speed up the solubilization process.
 68. The method of preparing the composition for solubilization of paclitaxel including emulsifier according to claim 66, wherein the said mixture is heated to 50° C. in step (2) to speed up the solubilization process.
 69. The method of preparing the composition for solubilization of paclitaxel including emulsifier according to claim 66 wherein the said mixture is sonicated in a bath type sonicator in step (2) to speed up the solubilization process.
 70. A method of preparing the composition for solubilization of paclitaxel including emulsifier according to any one of claims 33, wherein said method comprises the steps of: (1) preparing the paclitaxel solution by solubilizing 0.01˜20% by weight of paclitaxel in 0.01˜90% by weight of an oil chosen from triglyceride, iodized oil, vegetable oil and animal oil by sonicating in a bath type sonicator (step 1); and (2) preparing homogeneous mixture by mixing the paclitaxel solution in step (1) and 0.01˜90% by weight of at least one emulsifier and 4˜90% by weight of monoolein so that the ratio of monoolein to oil is more than 1:1 (step 2).
 71. The method of preparing the composition for solubilization of paclitaxel including emulsifier according to claim 70, wherein the said mixture is heated to 50° C. and sonicated in a bath type sonicator in step (2) to speed up the solubilization process.
 72. The composition for solubilization of paclitaxel including emulsifier according to claims 1, wherein the said composition is liquid or semi-solid state at room temperature.
 73. The composition for solubilization of paclitaxel according to claim 1, comprising 41.5˜66% by weight of monoolein, 27˜41.5% by weight of an oil selected from a group consisting of chosen from triglyceride, iodized oil, vegetable oil and animal oil and 0.4˜3% by weight of paclitaxel.
 74. The composition for solubilization of paclitaxel including emulsifier according to claim 33, wherein the said composition is liquid or semi-solid state at room temperature. 